Effect of Dimethyl Fumarate (DMF) on T-cell Acute Lymphoblastic Leukemia / 中国实验血液学杂志

OBJECTIVE@#To explore the effect and possible mechanism of dimethyl fumarate (DMF) on T-cell acute lymphoblastic leukemia (T-ALL), and provide experimental and theoretical basis for the clinical treatment of T-ALL.@*METHODS@#Jurkat cells were treated with different concentrations of DMF for 24 hours, and then the proportion and absolute count of Ki67-positive Jurkat cells were analyzed by flow cytometry. Meanwhile, the protein levels of nuclear factor-erythroid 2-related factor 2 (Nrf2) and E3 ubiquitin ligase HACE1 in Jurkat cells treated with DMF for 24 hours were evaluated by Western blot. Nrf2 proteins were co-immunoprecipitated in Jurkat cells, and then HACE1 protein was assessed by Western blot. Plasmids of Flag-Nrf2 and different gradients of Flag-HACE1 were transfected into HEK293T cells, and the levels of Flag-Nrf2 were detected by Western blot after 48 hours.@*RESULTS@#DMF could significantly inhibit the proportion and absolute count of Ki67-positive Jurkat cells, and DMF inhibited the proliferation of Jurkat cells in a dose-dependent manner (r=0.9595, r=0.9054). DMF could significantly up-regulate the protein levels of Nrf2 and E3 ubiquitin ligase HACE1 in Jurkat cells (P<0.01, P<0.01). HACE1 physically interacted with Nrf2 in Jurkat cells. Overexpression of Flag-HACE1 significantly increased the protein level of Flag-Nrf2 in a dose-dependent manner (r=0.9771).@*CONCLUSION@#DMF inhibits the proliferation of T-cell acute lymphoblastic leukemia cell. The mechanism may be that, DMF significantly up-regulates the protein levels of Nrf2 and E3 ubiquitin ligase HACE1, and HACE1 interacts with Nrf2 and positively regulates Nrf2 protein level.

Preliminary study on cerebrospinal fluid proteomics of Erxian Decoction against neurogenesis impairment in late-onset depression / 中国中药杂志

This study aims to elucidate the underlying mechanism of Erxian Decoction(EXD) against neurogenesis impairment in late-onset depression(LOD) rats based on cerebrospinal fluid(CSF) proteomics. A total of 66 20-21-month-old male Wistar rats were randomized into naturally aged(AGED) group, LOD group, and EXD group. All rats received chronic unpredictable mild stress(CUMS) for 6 weeks for LOD modeling except for the AGED group. During the modeling, EXD group was given EXD(ig, twice a day at 4 g·kg~(-1)) and other groups received equivalent amount of normal saline(ig). After modeling, a series of behavioral tests, such as sucrose preference test(SPT), open-field test(OFT), forced swimming test(FST), and Morris water maze test(MWMT) were performed. Immunofluorescence method was used to detect the number of Ki-67/Nesti-positive cells and BrdU/DCX-positive cells in the hippocampal DG area of each group. High-concentration corticosterone(CORT) was combined with D-galactose(D-gal) to simulate the changes of LOD-related stress and aging and the proliferation and differentiation of primary neural stem cells of hippocampus in each group were observed. Data independent acquisition(DIA)-mass spectrometry(MS) was used to analyze the differential proteins in CSF among groups and bioinformatics analysis was performed to explore the biological functions of the proteins. Behavioral tests showed that sucrose consumption in SPT, total traveling distance in OFT, and times of crossing the platform in MWMT were all reduced(P<0.01) and the immobility time in FST was prolonged(P<0.01) in the LOD group compared with those in the AGED group, suggesting that LOD rats had developed depression symptoms such as anhedonia, decreased locomotor activity ability, and cognitive dysfunction. Behavioral abnormalities were alleviated(P<0.01, P<0.05) in the EXD group as compared with those in the LOD group. Immunofluorescence results demonstrated that Ki-67/Nesti-positive cells and BrdU/DCX-positive cells in the hippocampal DG area were fewer(P<0.05) in LOD group than in the AGED group, and the positive cells in the EXD group were more(P<0.05) than those in the LOD group. In vitro experiment showed that the proliferation and differentiation of primary hippocampal neural stem cells under the CORT+D-gal treatment were reduced(P<0.01). The proliferation rate of neural stem cells decreased(P<0.05) in CORT+D-gal+LOD-CSF group but increased(P<0.01) in CORT+D-gal+EXD-CSF group compared with that in the CORT+D-gal group. A total of 2 620 proteins were identified from rat CSF, with 135 differential proteins between the LOD group and AGED group and 176 between EXD group and LOD group. GDF11, NrCAM, NTRK2, and GhR were related to neurogenesis and 39 differential proteins were regulated by both LOD and EXD. EXD demonstrated obvious anti-LOD effect, as it improved the locomotor activity ability and cognitive function of LOD rats and protected the proliferation and differentiation of hippocampal neural stem cells. EXD exerts anti-LOD effect by regulating the proteins related to neurogenesis in CSF, such as GDF11, NrCAM, NTRK2, and GhR and maintaining hippocampal neurogenesis.

Curcumin inhibited hypoxia induced epithelial-mesenchymal transition in hepatic carcinoma cell line HepG2 in vitro / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore effects and possible mechanisms of curcumin on hypoxia induced epithelial-mesenchymal transition (EMT) in hepatocellular carcinoma cell line HepG2.</p><p><b>METHODS</b>HepG2 cells were divided to 3 groups, i.e., the normal control group, the CoCl2 group, and the CoCl2 plus 10 micromol/L curcumin group. The proliferation of HepG2 was determined using MTT assay. The migration of HepG2 was detected by wound healing assay.The mRNA expression of hypoxia-inducible factor-1 (HIF-1alpha) was evaluated with real-time RT-PCR. The protein expressions of HIF-1alpha, epithelial-cadherin (E-cadherin), and vimentin were determined using Western blot.</p><p><b>RESULTS</b>Compared with the normal control group, the proliferation and migration of HepG2 cells under CoCl2-induced hypoxia significantly increased, the expression of HIF-1alpha was up-regulated, and the expression of E-cadherin protein was obviously down-regulated, and the expression of vimentin significantly increased (all P < 0.05). Intervention by curcumin significantly inhibited the proliferation and migration of hypoxic HepG2 cells, and expressions of HIF-1alpha and vimentin decreased, and the expression of E-cadherin was up-regulated, showing statistical difference when compared with those of the CoCl2 group (P < 0.05). There was no statistical difference in HIF-1alpha mRNA expression among the 3 groups (P > 0.05).</p><p><b>CONCLUSION</b>Curcumin could reverse the proliferation and migration of HepG2 cells under CoCl2-induced hypoxia condition, which might be associated with inhibiting up-regulated expressions of HIF-1alpha protein and EMT.</p>

Efficacy and safety of low-dose rituximab combined with different dosage of glucocorticoids for immune thrombocytopenia / 中华血液学杂志

<p><b>OBJECTIVE</b>To compare the efficacy and safety of low-dose rituximab combined with different dosage of glucocorticoids for immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Seventy-four patients (35 male, median age 34 years, range 18-70 years) including 60 newly-diagnosed, 6 persistent, 5 chronic and 3 refractory patients were enrolled in this study, and separated into control (36 cases) and experimental (38 cases) groups according to the dosage of glucocorticoids. Patients in both groups received dexamethasone 40 mg/day on days 1-4, followed by rituximab 100 mg on days 7, 14, 21, 28. The patients in experimental group also received decrements of prednisone 60, 30, 20, 10 mg/day on days 5-7, 8-14, 15-21, 22-28. The initial, long-term efficacy and safety were evaluated.</p><p><b>RESULTS</b>Platelet counts of all patients at day 4 remarkably increased, with the median platelet count from 11(1-26) × 10⁹/L to 84(23-132) × 10⁹/L in control group, and 10(2-20) × 10⁹/L to 80(22-115) × 10⁹/L in experimental group; the platelet counts of patients at day 14 in experimental group [163(19-262) × 10⁹/L] was higher than that of control group [98(18-238) × 10⁹/L] (P<0.05). The overall response (OR) rates at day 28 in experimental group (84.21%) was significantly higher than that of control group (66.67%, P = 0.03). There was no significant difference of sustained response (SR) rates in two groups (63.89%vs 65.79%, 58.33%vs 60.53%, P > 0.05) at six and twelve months follow-up points. Both groups showed similar incidence of adverse events, and no patients discontinued the treatment due to side effects.</p><p><b>CONCLUSION</b>Low-dose rituximab and glucocorticoids was an effective method for ITP patients, and maintenance treatment with decrements of prednisone contributed to improving earlier CR rate.</p>

Effect of different therapeutic regimens on regulatory T cells in patients of primary immune thrombocytopenia / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the change of CD4⁺CD25(high)CD127(low) regulatory T cells (Tregs) percentage in patients with primary immune thrombocytopenia (ITP) treated by different methods.</p><p><b>METHODS</b>One hundred and thirty-eight newly diagnosed adult ITP patients (57 male, median age 40 years, range 18-70 years) were enrolled in this study, who were randomly separated into three regiment groups, namely prednisolone (PSL, 1.5 mg/kg for 2-4 weeks and subsequently stepwise reduction) group enrolled 49 patients, dexamethasone [(one course of high-dose dexamethasone (HDD) 40 mg/day, d1-4] 45 patients, and rituximab plus HDD (rituximab 100 mg on days 7, 14, 21, 28 and HDD) group 44 patients. Peripheral blood was taken in ITP patients of each group before treatment, 14 d and 28 d after treatment. The percentages of CD4⁺CD25(high)CD127(low) Tregs in 30 healthy controls and 138 patients were analyzed by flow cytometry.</p><p><b>RESULTS</b>Overall response (OR) rates of PSL, HDD and R+HDD groups at day 28 were 69.4%, 66.7% and 79.5% respectively with no difference. After the following 12 months, sustained response (SR) was more pronounced in R+HDD group compared to the other two groups (R+HDD vs PSL: 66.7% vs 37.8%, P<0.05; R+HDD vs HDD: 66.7% vs 22.7%, P<0.05). The percentage of CD4⁺CD25(high)CD127(low) Tregs in peripheral blood of ITP patients [(1.67±0.70)%] was significantly lower than in healthy control group; After treatment, the percentages of Tregs in peripheral blood of patients both at day 14 and 28 in R+HDD group remarkably decreased compared with before treatment [(4.28±1.09)% vs (1.68±0.68)%, P<0.05; (4.44±0.63)% vs (1.68±0.68)%]. The percentages of Tregs at day 14 in both other two groups decreased notably compared with before treatment. But the Tregs levels measured at day 28 in PSL and HDD groups were similar with before treatment.</p><p><b>CONCLUSION</b>The percentage of CD4⁺CD25(high)CD127(low) Tregs in peripheral blood of ITP patients was lower than healthy individual. The higher SR of patients treated by R+HDD was related to its ability to up-regulate the percentage of CD4⁺CD25(high)CD127(low) Tregs.</p>

The expression of hepatitis B virus X protein and cyclooxygenase-2 in hepatitis B virus-related hepatocellular carcinoma: correlation with microangiogenesis and metastasis, and what is the possible mechanism / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expression of HBx and COX-2 in hepatitis B virus-related hepatocellular carcinoma, and Its correlation with microangiogenesis and metastasis, and possible mechanism.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expression of hepatitis B virus X, cyclooxygenase-2 and CD34 in hepatitis B virus related hepatic carcinoma and 22 non-HBV related hepatic carcinoma tissues. The expression of hepatitis B virus x protein and cyclooxygenase-2 in hepatitis B virus-related hepatocellular carcinoma correlated with microangiogenesis and metastasis was tested by Spearman correlation analysis. The expression of COX-2 in HepG2-X was detected by Western blot and RT-PCR. PGE2 was detected by ELISA in clear supernatant liquid of HepG2-X. Trypan blue exclusion was performed to examine the inhibitory effects of COX-2 selective inhibitor (celecoxib).</p><p><b>RESULTS</b>In Hepatitis B carcinoma tissue, HBx and COX-2 were expressed at high level. The positive rate of COX-2 expression was 88.87% (55/62) in HBx positive expression group, which was significantly higher than that of the positive expression 31.82% (7/22, x2=27.188, P<0.01) in HBx negative expression group and 40.91% (9/22, x2=20.453, P<0.01) in non-HBV related hepatic carcinoma tissues, but it had no statistical difference (x2=0.393, P=0.531) between the HBx negative expression group and non-HBV related hepatic carcinoma tissue group. The expressions of HBx and COX-2 in metastasis group were higher than that in non-metastasis group (P<0.01), MVD in HBx or COX-2 positive expression group was significantly higher than that in negative expression group and non-HBV related hepatic carcinoma tissues (P is less than 0.01). MVD with metastasis was higher than that without metastasis (P<0.01) and MVD with portal vein invasion was higher than that without invasion (P<0.05). Spearman correlation analysis showed that the expression of COX-2 was significantly correlated with the expression of HBx (Rs=0.568, P<0.01). Celecoxib suppressed the growth of both cells in a dose-dependent manner. HepG2-X was significantly susceptible to celecoxib as compared to the HepG2-PC cells. COX-2 protein and mRNA were upregulated in HepG2-X cells than in HepG2-PC. Moreover, PGE2 was upregulated in clear supernatant liquid of HepG2-X than in HepG2-PC.</p><p><b>CONCLUSION</b>The expressions of HBx and COX-2 were higher in HBV-related hepatocellular carcinoma. COX-2 was significantly correlated with HBx in HCC and it could be a key factor involved in HBx contributed hepatocellular carcinoma's microangiogenesis and metastasis. The possible mechanism of the dual effects might be through HBx, COX-2 and PGE2 pathways in infiltration involved metastasis and microangiogenesis involved metastasis.</p>

Expressions of hepatitis B virus x protein and hypoxia-inducible factor-1alpha in hepatocellular carcinoma and their possible relationships / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the expressions of hepatitis B virus x protein (HBx) and hypoxia-inducible factor-1alpha (HIF-1alpha) in hepatocellular carcinoma (HCC) tissues and HepG2 cells under normoxic and hypoxic conditions.</p><p><b>METHODS</b>Immunohistochemistry was used to detect the expressions of HBx and HIF-1alpha in 78 hepatic carcinoma tissues, and their possible relationships were analyzed using SPSS 10.0 statistical software. Immunofluorescence and Western blot were used to investigate the expression of HIF-1alpha in HepG2 and HepG2-X cells under normoxic and hypoxic conditions. Flow cytometric analysis was used to measure reactive oxygen species (ROS) in HepG2 and HepG2-Xd cells under normoxic and hypoxic conditions.</p><p><b>RESULTS</b>The positive rate of HBx in the hepatocellular carcinoma tissues was 74.36% (58/78), while the positive rate of HIF-1alpha was 69.23% (54/78). Under normoxic condition, HIF-1alpha was mainly localized in the cytoplasm and partially translocated into the nuclei of most HepG2-X cells, while under hypoxic condition the expression of HIF-1alpha was positive in the cytoplasm and nuclei of HepG2 and HepG2-X cells. Under normoxic condition the expression of HIF-1alpha was negative in the HepG2 cells while it was positive in HepG2-X cells, but HIF-1alpha was upregulated time-dependently in both HepG2 and HepG2-X cells under hypoxia condition. Furthermore, HepG2-X cells had a higher level of ROS than HepG2 cells under normoxic condition, while under hypoxic condition, the levels of ROS in HepG2 and HepG2-X cells were not significantly different, but the levels of ROS in HepG2 and HepG2-X cells under hypoxia condition were higher than those under normoxia condition.</p><p><b>CONCLUSION</b>HBx and HIF-1alpha were widely expressed in HCC tissues and there is a positive relationship between them. HIF-1alpha can be upregulated by HBx in HepG2 cells under normoxic and hypoxia conditions. Regulation of HIF-1alpha by HBx in HCC might be via the ROS pathway.</p>